Search Results (98)

Background: Distinguishing between osteoarthritis (OA), rheumatoid arthritis (RA), and psoriatic arthritis (PsA) remains challenging despite different underlying mechanisms. Synovial fluid reflects metabolic changes within affected joints, yet comprehensive metabolomic comparisons across these conditions are limited. We aimed to identify disease-specific metabolic signatures in synovial fluid that could improve differential diagnosis and reveal therapeutic targets. Methods: We collected synovial fluid from 39 patients (20 OA, 5 RA, and 14 PsA) during routine knee arthrocentesis between January 2023 and February 2024. Following metabolite extraction, we performed untargeted metabolomic profiling using quadrupole time-of-flight liquid chromatography–mass spectrometry (Q-TOF LC/MS). Data underwent multivariate statistical analysis, including principal component analysis (PCA) and partial least squares–discriminant analysis (PLS-DA), to identify discriminatory metabolites. Results: While unsupervised analysis showed overlap between groups, supervised PLS-DA achieved clear metabolic separation. RA samples showed elevated itaconic acid, indicating inflammatory macrophage activation, and increased O-acetylserine, suggesting altered one-carbon metabolism. Hypoxanthine was decreased, which reflected severe metabolic stress. PsA exhibited the unique elevation of 4,4-dimethylcholestane and 2-oxoarginine. These metabolites have previously been unreported in this disease. OA demonstrated increased hippuric acid and indoleacetic acid, which are both gut microbiota products, supporting the gut–joint axis hypothesis. Conclusions: Each arthritis type displayed distinct metabolic fingerprints in synovial fluid. Candidate discriminatory metabolites, including gut-derived metabolites in OA and specific lipid alterations in PsA, open new diagnostic and therapeutic avenues. Given the limited RA sample size (n = 5), RA-related results should be viewed as exploratory and requiring validation in larger independent cohorts. These metabolites may, after rigorous validation in larger and independent cohorts, contribute to multi-metabolite biomarker panels for earlier diagnosis and to the rational design of targeted therapeutics addressing disease-specific metabolic disruptions. Full article

Recently, some cases of intentional animal poisoning using Carbofuran (CF) occurred in the Czech Republic, although CF is no longer available in the EU market. The present study describes a novel non-targeted analysis (NTA) workflow developed to possibly characterize 13 CF formulations from various sources to be certainly identified in real samples. Furthermore, a detection and quantification method for CF was developed for analyzing three animal samples, obtained from dead animals. The analyses were conducted using the liquid electron ionization (LEI) interface coupled with a quadrupole time-of-flight (QTOF) mass spectrometer, allowing the simultaneous characterization of the formulation’s volatile and low-volatile fractions. Almost all compounds detected in the different formulations were identified by comparing the experimental spectra with the NIST library at high probability values (95–99.38%). Determination of molecular ions, followed by MS/MS analysis, was performed to confirm compound identities at a high level of confidence. The quantification method for CF was successfully validated, showing negligible matrix effects (107%). CF was detected in two out of the three real samples. Only 3-keto-carbofuran was detected in one of the real samples; without any other marker, it was not possible to identify the specific formulation used in the three poisoning cases. Full article

Objectives: This study systematically compared the differences in egg quality between the BIAN chicken, an indigenous breed of Shanxi Province, and the Hy-Line Brown, a commercial breed, through the integration of non-targeted metabolomics and volatile flavoromics methods. Methods: A total of 675 metabolites and 84 volatile flavor compounds were identified in eggs from 300-day-old laying hens using LC-MS and GC × GC-TOF MS techniques. Results: BIAN chicken eggs exhibited notable advantages in flavor quality. The relative odor activity value (ROAV) of 1-octen-3-ol, a key flavor component, was 27.01 in BIAN compared with 13.46 in Hy-Line Brown, contributing to the characteristic mushroom aroma of BIAN eggs. Furthermore, the levels of heptaldehyde, 2-pentylfuran, and styrene in BIAN chicken eggs were significantly elevated, contributing to its characteristic flavor profile. Metabolomic analysis identified 40 breed-specific metabolites in BIAN chicken, with 21 up-regulated and 19 down-regulated. These metabolites were primarily involved in biological processes such as α-linolenic acid metabolism, cholesterol metabolism, and unsaturated fatty acid biosynthesis, highlighting the distinctive lipid metabolism regulation in BIAN chicken. Sensory evaluation based on relative odor activity values (ROAV) demonstrated that BIAN chicken eggs exhibited enhanced sweet, fruity, herbal, and citrus aromas, which correlated with the enriched lipid metabolism pathways. Conclusions: This study elucidates the molecular basis of distinctive egg quality characteristics in local chicken breeds, offering a scientific rationale for the conservation and utilization of indigenous breeds and the documentation of their unique metabolic and sensory properties. Furthermore, it furnishes a theoretical framework for understanding breed-specific flavor development and provides baseline data for future genetic selection and nutritional intervention strategies. Full article

Background: Platelet-rich plasma (PRP) is a widely used therapeutic product in musculoskeletal treatments due to its regenerative and anti-inflammatory properties. However, the lack of standardization in PRP preparation protocols hampers clinical consistency. Methods: In this study, the metabolic profiles of 10 different PRP types were compared using untargeted metabolomics via Q-TOF LC–MS. PRP-G and PRP-S were prepared from six donors to assess inter-individual variability, while the remaining types were obtained from a single donor to isolate the impact of preparation method alone. Multivariate analyses, VIP scores, and pathway enrichment analyses were conducted. Results: PRP formulations exhibited distinct metabolic differences associated with inflammatory signaling, redox homeostasis, steroid metabolism, energy production, and platelet activation. Samples from both single- and multi-donor groups showed high intra-group similarity, indicating that preparation method is a major determinant of PRP’s biochemical composition. Conclusion: Metabolomic profiling reveals that even minor differences in PRP preparation protocols can lead to significant biochemical changes that may affect therapeutic outcomes. This study highlights the need for standardized, indication-specific PRP products and underscores the value of metabolomic analysis in guiding optimal formulation selection in clinical practice. Full article

Scutellaria species are widely utilized and have demonstrated diverse biological effects for various diseases, both globally and in traditional Chinese medicine, due to the presence of bioactive compounds with unique structures. This study was conducted to reveal the in vitro effects and phytochemical properties of Scutellaria salviifolia Benth., an endemic species of Türkiye. The inhibitory effects of methanol extracts prepared separately from the aerial and root parts of S. salviifolia on the COX-2 and 5-LOX enzymes and their DPPH and ABTS radical scavenging activities were evaluated using in vitro methods. Additionally, the phenolic compounds of the extracts were compared based on Q-TOF LC/MS analysis. The extracts of S. salviifolia exhibited a high inhibitory effect on COX-2 enzyme activity, comparable to that of celecoxib. Still, they showed no significant effects in the 5-LOX enzyme inhibition assay. In the antioxidant activity assays, the percentage of inhibitory effects of both extracts against DPPH and ABTS were similar. A total of 29 and 27 compounds were detected in the aerial part and root extracts, respectively. Among the identified compounds, 18 were common to both the aerial part and root extracts. S. salviifolia may serve as a valuable alternative to the most well-known Scutellaria species, including S. baicalensis and S. barbata. Full article

Most studies on the Arthrosphaera genus, or giant pill millipedes, focus on its taxonomy, distribution, and ecology. Therefore, this investigation aimed to explore the chemical composition and bioactive properties of the hemolymph of the giant pill millipede Arthrosphaera lutescens (Butler, 1872). Chemical characterization of hemolymph was performed using gas chromatography–mass spectrometry (GC-MS) and liquid chromatography–quadrupole time-of-flight mass spectrometry (LC-MS Q-TOF), revealing a complex array of over 200 compounds. The bioactive properties of hemolymph were determined by using radical scavenging capacity (DPPH assay); antibacterial activity against human pathogens like Escherichia coli (Migula, 1895) Castellani and Chalmers 1919, Klebsiella pneumonia (Schroeter, 1886) Trevisan 1887, and Staphylococcus aureus (Rosenbach, 1884); and cytotoxicity against Dalton’s lymphoma ascites (DLA) cells using the trypan blue assay. The hemolymph showed radical scavenging properties and antibacterial and cytotoxic activity. Among the identified metabolites, 1,2-dimethoxy-13-methyl-[1,3]benzodioxolo[5,6-c]phenanthridine (DMBP) emerged as a promising candidate due to its high abundance and bioactivity profile, showcasing therapeutic potential against both lymphoma and S. aureus in further docking studies. Computational analysis identified key T-cell lymphoma targets, with molecular docking suggesting DMBP’s anticancer properties through interactions with proteins like AKT1 and mTOR. Additionally, docking revealed DMBP’s antibacterial effects via interactions with proteins such as Sortase-A and DNA gyrase. This research underscores the potential pharmaceutical applications of metabolites from giant pill millipedes. Full article

Vitamin E deficiency (VED) represents a common micronutrient deficiency in dairy cows (DCs), leading to severe degenerative diseases, oxidative stress, immune dysfunction, and various health issues, ultimately causing significant economic losses for the global dairy sector. Accordingly, our objective was to explore the metabolic features of VED-afflicted cows by combining the untargeted gas chromatography-time-of-flight mass spectrometry (GC-TOF-MS) and targeted liquid chromatography-mass spectrometry (LC-MS) to identify effective serum VED biomarkers. Untargeted GC-TOF-MS analysis identified 31 differential metabolites (DMs): 20 were overexpressed and 11 were suppressed in the VED group compared to the healthy control group. These DMs were enriched in six major metabolic pathways: glycine, serine, and threonine; alanine, aspartate, and glutamate; cysteine and methionine; tyrosine; primary bile acid biosynthesis; and nitrogen metabolisms. These outcomes show that VED significantly disrupts amino acid/lipid/energy metabolism pathways in DCs. Further targeted LC-MS quantification revealed significant alterations in key metabolites, including increased levels of norepinephrine, glycine, cysteine, and L-glutamine, as well as a significant reduction in cholesterol concentrations. Binary logistic regression analysis identified norepinephrine and cholesterol as strong candidate biomarkers for VED. Receiver operating characteristic curve analysis established outstanding diagnostic accuracy for norepinephrine and cholesterol (for both p < 0.001, area under the curve = 0.980 and 0.990, correspondingly), with sensitivities and specificities of 90% and 100%, respectively. In conclusion, this study integrates untargeted and targeted metabolomics approaches to reveal VED-caused metabolic disruptions in DCs, particularly in amino acid/lipid/energy metabolism pathways. Norepinephrine and cholesterol were identified as highly accurate serum VED biomarkers with excellent diagnostic performance. Early detection and timely intervention using these biomarkers could promote disease treatment and cow health, as well as productivity, and decrease economic losses. Full article

Glioblastoma is the most common and aggressive brain tumor in adults. However, due to the limitations of conventional treatments, as well as their side effects, there is a need to develop more effective and less harmful therapy strategies. There is evidence that plants endemic to northern Mexico possess biological activities that positively impact human health, particularly against cancer. Echinocereus engelmannii and Echinocereus pectinatus are cacti from the north of Mexico that produce bioactive compounds with antitumor activity. We obtained methanol extracts from previously isolated and fermented microorganisms associated with these cacti. Cell lines of extracts with cytotoxicity against glioblastoma cells U87, neuroblastoma cells SH-S5Y5, and Schwann neuronal cells (healthy control) were evaluated, using a colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazole bromide (MTT) reduction technique. The selective cytotoxicity extracts were analyzed using liquid chromatography tandem mass spectrometry (LC/MS²). We isolated 19 endophytic and soil-associated microorganisms from both cacti. Two of them were selected for their high percentages of tumor growth inhibition. The microorganism ES4 possessed the best activity with an IC₅₀ of 17.31 ± 1.70 µg/mL and a selectivity index of 3.11. We identified the bacterium Stenotrophomonas maltophilia by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) from the most active microorganisms against tumor growth. LC/MS² characterized the HS4 extract, and the most abundant group (50.0%) identified included carboxylic acids and derivatives, particularly bisgerayafolin A, Cyclo (Pro-Leu), maculosin, and tryptophan. In conclusion S. maltophilia extract inhibit the growth of glioma cells, showing greater sensitivity in the U87 cell line. Full article

Melittin, a naturally occurring antimicrobial peptide, demonstrates broad-spectrum activity, effectively suppressing and eliminating both Gram-positive and Gram-negative bacteria, including specific drug-resistant strains. In this study, molecular simulation software was employed to investigate and modify the structure of melittin with the aim of synthesizing a modified peptide exhibiting enhanced antibacterial potency and assessing its bacteriostatic and antibacterial properties. The primary research objectives were as follows: 1. Preparation and characterization of melittin-modified peptide—Using molecular simulation software, the structure of the melittin-modified peptide was adjusted to predict its activity and select the most appropriate amino acid sequence. The peptide was synthesized through solid-phase peptide synthesis employing the Fmoc strategy and subsequently purified using liquid chromatography. The yield of the purified modified melittin was determined to be 30.97%, and the identity of the product was confirmed by LC-MS and MALDI-TOF-MS. 2. Evaluation of the antimicrobial activity of the melittin-modified peptide—The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of melittin and its modified peptide were measured using gradient dilution and plate counting techniques. The results revealed that both melittin and its modified peptide exhibited strong antibacterial efficacy against Gram-positive and Gram-negative bacteria, as well as certain drug-resistant strains. This showed that melittin and its modified peptide have the same antibacterial (killing) effect. A scanning electron microscope analysis indicated that both melittin and its modified peptide were capable of disrupting bacterial cell structures, leading to bacterial cell death. Full article

Background/Objectives: Inflammatory bowel disease (IBD) is a persistent inflammatory condition affecting the gastrointestinal tract, distinguished by the impairment of the intestinal epithelial barrier, dysregulation of the gut microbiota, and abnormal immune responses. Cajanus cajan (L.) Millsp., traditionally used in Chinese herbal medicine for gastrointestinal issues such as bleeding and dysentery, has garnered attention for its potential therapeutic benefits. However, its effects on IBD remain largely unexplored. Methods: In this study, the major compounds from Cajanus cajan leaf extract (CCLE) were initially characterized by LCMS-IT-TOF. The IBD model was developed in C57BL/6 mice by administering continuous 4% (w/v) dextran sodium sulfate (DSS) aqueous solution over a period of seven days. The body weight, colon length, disease activity index (DAI), and histopathological examination using hematoxylin and eosin (H&E) staining were performed in the IBD model. The levels of the main inflammatory factors, specifically TNF-α, IL-1β, IL-6, and myeloperoxidase (MPO), were quantified by employing enzyme-linked immunosorbent assay (ELISA) kits. Additionally, the levels of tight junction proteins (ZO-1, Occludin) and oxidative stress enzymes (iNOS, SOD1, CAT) were investigated by qPCR. Subsequently, flow cytometry was employed to analyze the populations of various immune cells within the spleen, thereby assessing the impact of the CCLE on the systemic immune homeostasis of IBD mice. Finally, 16S rDNA sequencing was conducted to examine the composition and relative abundance of gut microbiota across different experimental groups. In addition, molecular docking analysis was performed to assess the interaction between the principal components of CCLE and the aryl hydrocarbon receptor (AHR). Results: We identified seven bioactive compounds in CCLE: catechin, cajachalcone, 2-hydroxy-4-methoxy-6-(2-phenylcinyl)-benzoic acid, longistylin A, longistylin C, pinostrobin, amorfrutin A, and cajaninstilbene acid. Our results demonstrated that oral administration of CCLE significantly alleviates gastrointestinal symptoms in DSS-induced IBD mice by modulating the balance of gut-derived pro- and anti-inflammatory cytokines. This modulation is associated with a functional correction in M1/M2 macrophage polarization and the Th17/Treg cell balance in splenic immune cells, as well as shifts in the populations of harmful bacteria (Erysipelatoclostridium and Staphylococcus) and beneficial bacteria (Odoribacter, unidentified Oscillospiraceae, Lachnoclostridium, and Oscillibacter) in the gut. Furthermore, cajaninstilbene acid, longistylin A, and longistylin C were identified as potential AhR agonists. Conclusions: The present results suggested that CCLE, comprising stilbenes like cajaninstilbene acid, longistylin A, and longistylin C, protects the epithelial barrier’s structure and function against DSS-induced acute IBD by restoring gut microbiota balance and systemic immune response as AhR agonists. Overall, CCLE represents a promising natural product-based therapeutic strategy for treating IBD by restoring gut microbiota balance and modulating systemic immune responses. Full article

A comprehensive LC-MS study examined the venom components of the solitary scoliid wasp Scolia oculata. Online mass fingerprinting showed that crude venom contains 25 small molecules (amino acids, biogenic amines, and nucleosides/nucleotides) and 45 peptides with MW 400-2700. The small molecules were identified by elemental composition analysis, and peptide sequences were determined by ESI-MS/MS and MALDI-TOF/TOF MS analyses. As major peptide components, a known peptide, β-scoliidine (DYVTVKGFSPLRKA), and three new peptides, γ-scoliidine (YVTVKGFSPLR), δ-scoliidine (YVTVKGFSPLREP) and ε-scoliidine (DYVTVKGFSPLREP) were identified, all of which are closely homologous to each other. Once the neuroprotective effects of β-scoliidine have already been described, the other three new scoliidine peptides were analyzed against oxidative stress-induced toxicity in PC12 neuronal cells by mitochondrial metabolism assay, and the structure-activity relationship was evaluated. Interestingly, pre-treatment with ε-scoliidine increased the mitochondrial metabolism of PC12 cells (106 ± 3.6%; p = 0.007) exposed to H₂O₂-induced oxidative stress in contrast to γ- and δ-scoliidines (77.6 ± 4.8 and 68.5 ± 4.1%, respectively) in compared to cells treated only H₂O₂ (75.8 ± 2.4%). These new peptides were also analyzed for enzyme inhibitor/substrate assays with angiotensin-converting enzyme (ACE), neprilysin (NEP), and acetylcholinesterase (AChE). In these assays, only δ- and ε-scoliidines increased the AChE activity (128.7 ± 3.8%; p = 0.01; and 116.8 ± 3.8% p = 0.03; respectively) in relation to basal activity (100.1 ± 1.6%). In addition, the four peptides were analyzed through in silico analysis, and none of them demonstrated possible hemolytic and toxic activities. In our study, the comprehensive LC-MS and MS/MS analyses of Scolia oculate venom identified four major peptide components of the venom β-, γ-, δ- and ε-scoliidines, and small differences in their primary structures are important to their neuroprotective properties. Full article

Phospholipids (PLs), essential components of cell membranes, play significant roles in maintaining the structural integrity and functionality of joint tissues. One of the main components of synovial joint fluid (SJF) is PLs. Structures such as PLs that are found in low amounts in biological fluids may need to be selectively enriched to be analyzed. Monodisperse-mesoporous SiO₂ microspheres were synthesized by a multi-step hydrolysis condensation method for the selective enrichment and separation of PLs in the SJF. The microspheres were characterized by SEM, XPS, XRD, and BET analyses. SiO₂ microspheres had a 161.5 m²/g surface area, 1.1 cm³/g pore volume, and 6.7 nm pore diameter, which were efficient in the enrichment of PLs in the SJF. The extracted PLs with sorbents were analyzed using Q-TOF LC/MS in a gradient elution mode with a C18 column [2.1 × 100 mm, 2.5 μM, Xbridge Waters (Milford, MA, USA)]. An untargeted lipidomic approach was performed, and the phospholipid enrichment was successfully carried out using the proposed solid-phase extraction (SPE) protocol. Recovery of the SPE extraction of PLs using sorbents was compared to the classical liquid–liquid extraction (LLE) procedure for lipid extraction. The results showed that monodisperse-mesoporous SiO₂ microspheres were eligible for selective enrichment of PLs in SJF samples. These microspheres can be used to identify PLs changes in articular joint cartilage (AJC) in physiological and pathological conditions including osteoarthritis (OA) research. Full article

The synthesis of silver nanoparticles (AgNPs) using environmentally friendly methods has become increasingly important due to its sustainability and cost-effectiveness. This study investigates the green synthesis of AgNPs using gall extracts from the plant Ficus recemosa, known for its high phytochemical content. The formation of AgNPs was verified through multiple analytical techniques, including UV-Vis spectroscopy, Fourier-transform infrared (FTIR) spectroscopy, transmission electron microscopy (TEM), zeta potential analysis, and dynamic light scattering (DLS). The UV-Vis spectroscopy results displayed a distinct surface plasmon resonance peak indicative of AgNP formation. FTIR analysis revealed specific interactions between silver ions and phytochemicals in the gall extract, while TEM images confirmed the nanoscale morphology and size of the synthesized particles. Zeta potential and DLS analyses provided insights into the stability and size distribution of the AgNPs, demonstrating good colloidal stability. Biological properties of the AgNPs were assessed through various assays. Antimicrobial activity was tested using the disc diffusion method against Escherichia coli and Staphylococcus aureus, showing significant inhibitory effects. The anticancer potential was evaluated using the trypan blue exclusion assay on Dalton’s Lymphoma Ascites (DLA) cells, revealing considerable cytotoxicity. Additionally, antimitotic activity was studied in the dividing root cells of Allium cepa, where the AgNPs significantly inhibited cell division. This research highlights the effective use of F. recemosa gall extracts for the green synthesis of AgNPs, presenting an eco-friendly approach to producing nanoparticles with strong antimicrobial, anticancer, and antimitotic properties. The promising results suggest potential applications of these biogenic AgNPs in medical and agricultural sectors, paving the way for further exploration and utilization. Full article

Onions are known not only for their culinary importance but also for their nutritional and health-promoting properties. Both properties are closely linked to their content of organosulfur compounds, which account for up to 5% of the dry weight of an onion. Given the importance of these compounds, suitable analytical methods are required for their study. Two techniques should be highlighted in this context: gas chromatography coupled to mass spectrometry (GC-MS), and liquid chromatography coupled to mass spectrometry (LC-MS). In this study, eight different onion varieties were analyzed using two distinct analytical techniques: direct thermal desorption–gas chromatography–mass spectrometry (DTD-GC-MS) and high-resolution mass spectrometry (HRMS) on an LC-ESI-QqTOF instrument. Each method identified different organosulfur compounds, with LC-HRMS targeting 15 non-volatile compounds, such as cysteine sulfoxides, and GC-MS targeting 18 volatiles, such as disulfides and trisulfides. The results obtained were studied using Pearson correlations and principal component analysis. No precise correlation was found between the initial organosulfur compounds in onions and their hydrolysates. Consequently, although GC is one of the most employed techniques in the scientific literature, the use of LC-HRMS or a combination of both techniques may offer a more comprehensive and accurate description of the metabolomic profile of onions. Full article

The main objective of this study was to investigate the metabolism of miconazole, an azole antifungal drug. Miconazole was subjected to incubation with human liver microsomes (HLM) to mimic phase I metabolism reactions for the first time. Employing a combination of an HLM assay and UHPLC-HRMS analysis enabled the identification of seven metabolites of miconazole, undescribed so far. Throughout the incubation with HLM, miconazole underwent biotransformation reactions including hydroxylation of the benzene ring and oxidation of the imidazole moiety, along with its subsequent degradation. Additionally, based on the obtained results, screen-printed electrodes (SPEs) were optimized to simulate the same biotransformation reactions, by the use of a simple, fast, and cheap electrochemical method. The potential toxicity of the identified metabolites was assessed using various in silico models. Full article